Team:Michigan/Oil Sands October
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- | Ben moved transformation plates to | + | Ben moved transformation plates to 4ºC yesterday, control plates were blank, and all plates except for J61100 appeared to have colonies. The Flu/pSB1AT3 ligation plates had very small colonies, so they were returned to the incubator with the J61100 plate. Also, it could be seen that many colonies on the 1:3 ligation plate appeared red, indicating that the vector had preferentially reformed. This could be because of the ratio, but dephosphorylation treatment should be used in the future for ligations. |
- | + | OD600s were taken of the Pseudomonas cultures (46 hours): | |
- | *Putida pH 7- 0.157 | + | *Putida pH 7 - 0.157 |
- | *Fluorescens pH 7- 0.129 | + | *Fluorescens pH 7 - 0.129 |
- | *Co-culture pH 7- 0.188 | + | *Co-culture pH 7 - 0.188 |
- | *Co-culture pH 9- 0.232 | + | *Co-culture pH 9 - 0.232 |
Surprisingly, they seem to grow better at pH 9, at least in co-culture. As expected, the co-culture did better than pure cultures. This lends some support towards Alex's biofilm assay results, however, it should still be repeated. Also, plated a serial dilution of the pH 9 co-culture, at 10<sup>-5</sup>, 10<sup>-6</sup>, and 10<sup>-7</sup> dilutions to determine the conversion factor between OD600 and cell count. Made streak plates of all other cultures, pure cultures for stock, and pH 7 co-culture to determine if colonies could be distinguished. All cultures and plates were returned to the 30<sup>o</sup>C incubator, without shaking. | Surprisingly, they seem to grow better at pH 9, at least in co-culture. As expected, the co-culture did better than pure cultures. This lends some support towards Alex's biofilm assay results, however, it should still be repeated. Also, plated a serial dilution of the pH 9 co-culture, at 10<sup>-5</sup>, 10<sup>-6</sup>, and 10<sup>-7</sup> dilutions to determine the conversion factor between OD600 and cell count. Made streak plates of all other cultures, pure cultures for stock, and pH 7 co-culture to determine if colonies could be distinguished. All cultures and plates were returned to the 30<sup>o</sup>C incubator, without shaking. |
Revision as of 00:59, 27 October 2010